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Mar. Drugs 2011, 9(9), 1487-1501; doi:10.3390/md9091487
Article

Characterization of a Novel Serine Protease Inhibitor Gene from a Marine Metagenome

1,†
,
1,†
,
1
,
1
,
1
 and
2,*
Received: 2 August 2011 / Revised: 22 August 2011 / Accepted: 25 August 2011 / Published: 5 September 2011
(This article belongs to the Special Issue Metagenomics in Biodiscovery from Oceans)
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Abstract

A novel serine protease inhibitor (serpin) gene designated as Spi1C was cloned via the sequenced-based screening of a metagenomic library from uncultured marine microorganisms. The gene had an open reading frame of 642 base pairs, and encoded a 214-amino acid polypeptide with a predicted molecular mass of about 28.7 kDa. The deduced amino acid sequence comparison and phylogenetic analysis indicated that Spi1C and some partial proteinase inhibitor I4 serpins were closely related. Functional characterization demonstrated that the recombinant Spi1C protein could inhibit a series of serine proteases. The Spi1C protein exhibited inhibitory activity against α-chymotrypsin and trypsin with Ki values of around 1.79 × 10−8 and 1.52 × 10−8 M, respectively. No inhibition activity was exhibited against elastase. Using H-D-Phe-Pip-Arg-pNA as the chromogenic substrate, the optimum pH and temperature of the inhibition activity against trypsin were 7.0–8.0 and 25 °C, respectively. The identification of a novel serpin gene underscores the potential of marine metagenome screening for novel biomolecules.
Keywords: serine protease inhibitor; uncultured marine microorganisms; sequenced-based screening; functional characterization serine protease inhibitor; uncultured marine microorganisms; sequenced-based screening; functional characterization
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Jiang, C.-J.; Hao, Z.-Y.; Zeng, R.; Shen, P.-H.; Li, J.-F.; Wu, B. Characterization of a Novel Serine Protease Inhibitor Gene from a Marine Metagenome. Mar. Drugs 2011, 9, 1487-1501.

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